Fluorescence labeling oligonucleotide probe and uses thereof

Abstract

The invention discloses a fluorescence marked obligonucleotide probe and an application of the probe. The probe of the invention changes the designing method of traditional Taqman probe that a fluorescence reporting group and a fluorescence quenching group are put at the two ends of the probe. The invention designs the fluorescence quenching group at the middle of the probe and meanwhile decorates two ends of the fluorescence quenching group with a fluorescence reporting group. Thereby, not only the distance between the fluorescence reporting group and the fluorescence quenching group is shortened, but the quenching effect is enhanced. And a molecular probe of the invention can release two molecular of fluorescence when being hydrolyzed, which can increase the strength of the fluorescence signal and enhance the detecting sensitivity. The invention can be widely applied to a real-time fluorescence PRC reaction and used for biological ship detecting.

Description

technical field [0001] The invention relates to a fluorescently labeled oligonucleotide probe and its application. Background technique [0002] Nucleic acid detection is becoming more and more important with its wide application in genotyping, genetic disease diagnosis, pathogenic microorganism detection and other fields, and the nucleic acid amplification technology based on polymerase chain reaction (PCR) provides these applications with A powerful tool [Chan YR, Morris A. Curr Opin Infect Dis. 2007; 20(2): 157-64; Malinowski DP. Expert Rev Mol Diagn. 2007; 7(3): 269-80.]. Moreover, with the emergence of real-time fluorescent PCR technology, nucleic acid detection no longer depends on gel electrophoresis detection after amplification, and becomes more convenient and practical [Parashar D. et al. Indian J Med Res. 2006; 124 (4 ):385-98; Valasek MA, Repa JJ. AdvPhysiol Educ. 2005;29(3):151-9.]. In the 1990s, the American Perkin Elmer (PE) company developed the Taqman fluo...

AI Technical Summary

Problems solved by technology

However, in actual use, when the probe is long, the distance between the fluorescent donor and acceptor groups at both ends of the probe is relatively long, resulting in a decrease in fluorescence quenching efficiency, and some quenching groups themselves will also generate fluorescent signals. , which will make the background of the reaction high and reduce the sensitivity of the detection

However, if the length of the probe is shortened in order to increase the quenching effect, the Tm value of the probe will be too low to affect the binding to the template, thereby reducing the detection efficiency.

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